Characterisation of peri-implantation endometrial Treg and identification of an altered phenotype in recurrent pregnancy loss

AbstractRecurrent Pregnancy Loss (RPL) affects 2-4% of couples, and with increasing numbers of pregnancy losses the risk of miscarrying a euploid pregnancy is increased, suggesting RPL is a pathology distinct from sporadic miscarriage that is due largely to lethal embryonic aneuploidy. There are a number of conditions associated with RPL including unspecified ‘immune’ pathologies, one of the strongest candidates for dysregulation remains T regulatory cells as depletion in the very early stages of pregnancy in mice leads to pregnancy loss.Human endometrial Treg and conventional CD4T cells were isolated during the peri-implantation period of the menstrual cycle in normal women. We identified an endometrial Treg transcriptomic signature and validated an enhanced regulatory phenotype compared to peripheral blood Treg. Parous women had an altered endometrial Treg transcriptome compared to nulliparity, indicating acquired immunity memory of pregnancy within the Treg population, by comparison endometrial conventional CD4T cells were not altered. We compared primary and secondary RPL to nulliparous or parous controls respectively. Both RPL subgroups displayed differentially expressed Treg gene transcriptomes compared to controls. We found increased cell surface S1PR1 and decreased TIGIT protein expression by Treg in primary RPL, confirming the presence of altered Treg in the peri-implantation RPL endometrium.

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Characterisation of peri-implantation endometrial Treg and identification of an altered phenotype in recurrent pregnancy loss

Mucosal Immunology ◽

10.1038/s41385-021-00451-1 ◽

2021 ◽

Author(s):

Ingrid Granne ◽

Mengni Shen ◽

Helena Rodriguez-Caro ◽

Gurmeher Chadha ◽

Elizabeth O’Donnell ◽

...

Keyword(s):

Pregnancy Loss ◽

Recurrent Pregnancy Loss ◽

T Regulatory Cells ◽

Immune Memory ◽

Regulatory Cells ◽

Treg Population ◽

Transcriptomic Signature ◽

Normal Women ◽

Implantation Period ◽

Regulatory Phenotype

AbstractRecurrent Pregnancy Loss (RPL) affects 2–4% of couples, and with increasing numbers of pregnancy losses the risk of miscarrying a euploid pregnancy is increased, suggesting RPL is a pathology distinct from sporadic miscarriage that is due largely to lethal embryonic aneuploidy. There are a number of conditions associated with RPL including unspecified “immune” pathologies; one of the strongest candidates for dysregulation remains T regulatory cells as depletion in the very early stages of pregnancy in mice leads to pregnancy loss. Human endometrial Treg and conventional CD4T cells were isolated during the peri-implantation period of the menstrual cycle in normal women. We identified an endometrial Treg transcriptomic signature and validated an enhanced regulatory phenotype compared to peripheral blood Treg. Parous women had an altered endometrial Treg transcriptome compared to nulliparity, indicating acquired immune memory of pregnancy within the Treg population, by comparison endometrial conventional CD4T cells were not altered. We compared primary and secondary RPL to nulliparous or parous controls respectively. Both RPL subgroups displayed differentially expressed Treg gene transcriptomes compared to controls. We found increased cell surface S1PR1 and decreased TIGIT protein expression by Treg in primary RPL, confirming the presence of altered Treg in the peri-implantation RPL endometrium.

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Faculty Opinions recommendation of GRAIL is up-regulated in CD4+ CD25+ T regulatory cells and is sufficient for conversion of T cells to a regulatory phenotype.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1063800.516726 ◽

2007 ◽

Author(s):

Stephen Cobbold

Keyword(s):

T Cells ◽

T Regulatory Cells ◽

Regulatory Cells ◽

Regulatory Phenotype

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Multiple sclerosis patients have reduced resting and increased activated CD4+CD25+FOXP3+T regulatory cells

Scientific Reports ◽

10.1038/s41598-021-88448-5 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Nirupama D. Verma ◽

Andrew D. Lam ◽

Christopher Chiu ◽

Giang T. Tran ◽

Bruce M. Hall ◽

...

Keyword(s):

T Cells ◽

T Regulatory Cells ◽

Mononuclear Cells ◽

Effector Memory ◽

Regulatory Cells ◽

Peripheral Blood Mononuclear ◽

Healthy Donors ◽

Treg Population ◽

Multiple Sclerosis Patients ◽

Population Iii

AbstractResting and activated subpopulations of CD4+CD25+CD127loT regulatory cells (Treg) and CD4+CD25+CD127+ effector T cells in MS patients and in healthy individuals were compared. Peripheral blood mononuclear cells isolated using Ficoll Hypaque were stained with monoclonal antibodies and analysed by flow cytometer. CD45RA and Foxp3 expression within CD4+ cells and in CD4+CD25+CD127loT cells identified Population I; CD45RA+Foxp3+, Population II; CD45RA−Foxp3hi and Population III; CD45RA−Foxp3+ cells. Effector CD4+CD127+ T cells were subdivided into Population IV; memory /effector CD45RA− CD25−Foxp3− and Population V; effector naïve CD45RA+CD25−Foxp3−CCR7+ and terminally differentiated RA+ (TEMRA) effector memory cells. Chemokine receptor staining identified CXCR3+Th1-like Treg, CCR6+Th17-like Treg and CCR7+ resting Treg. Resting Treg (Population I) were reduced in MS patients, both in untreated and treated MS compared to healthy donors. Activated/memory Treg (Population II) were significantly increased in MS patients compared to healthy donors. Activated effector CD4+ (Population IV) were increased and the naïve/ TEMRA CD4+ (Population V) were decreased in MS compared to HD. Expression of CCR7 was mainly in Population I, whereas expression of CCR6 and CXCR3 was greatest in Populations II and intermediate in Population III. In MS, CCR6+Treg were lower in Population III. This study found MS is associated with significant shifts in CD4+T cells subpopulations. MS patients had lower resting CD4+CD25+CD45RA+CCR7+ Treg than healthy donors while activated CD4+CD25hiCD45RA−Foxp3hiTreg were increased in MS patients even before treatment. Some MS patients had reduced CCR6+Th17-like Treg, which may contribute to the activity of MS.

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Endogenous IL-β Selectively Converts T Regulatory Cells Into IL-17 Producers During Antigen Stimulation

Blood ◽

10.1182/blood.v116.21.586.586 ◽

2010 ◽

Vol 116 (21) ◽

pp. 586-586

Author(s):

Lequn Li ◽

Jin sub Kim ◽

Vassiliki A Boussiotis

Keyword(s):

T Cells ◽

T Regulatory Cells ◽

Neutralizing Antibody ◽

Treg Cells ◽

Conversion Process ◽

Regulatory Cells ◽

Antigen Stimulation ◽

Tnf Α ◽

Treg Population

Abstract Abstract 586 A major challenge of the immune system is to fight pathogens and tumor antigens while preserving tolerance to self-antigen. T regulatory cells (Treg) are critical extrinsic regulators of immune tolerance and maintenance of lymphoid homeostasis. Recently it was determined that, when used as cell-based immunosuppressive therapy, Treg have a potent effect in preventing GvHD in patients undergoing allogeneic stem cell transplantation. However, several studies suggest that the Treg phenotype is not at end stage of differentiation. Treg can express and produce effector cytokines including IFN-γ and IL-17 under certain conditions, particularly in the context of inflammatory milieu, suggesting that Treg may convert into inflammatory mediators. IL-1β and TNF-α are critical inflammatory cytokines that have been implicated in GvHD. The precise role and the mechanism(s) via which these cytokines may affect development of GvHD remain unclear. In the presence study, we sought to determine whether IL-1β and TNF-α regulate the properties of Treg and specifically whether these cytokines affect Treg expansion and/or conversion into IL-17 producing cells. CD4+CD25+Treg cells were isolated from B6 mice and were stimulated with anti-CD3-plus-anti-CD28 mAbs in the presence of either media, IL-1β or TNF-α. Addition of either cytokine induced Treg proliferation as determined by CFSE. Assessment of intracellular IL-17 expression by flow cytometry and IL-17 production by ELISA revealed that IL-1β but not TNF-α induced conversion of Treg into IL-17 producing cells, suggesting that conversion was mediated via pathways distinct from those that regulate cell cycle progression. To evaluate conversion of Treg to IL-17 producers during antigen stimulation and to determine the role of IL-1β in this process, we used neutral culture conditions in which no exogenous cytokines were supplied. Treg cells isolated from Foxp3GFP-KI mouse were added to cultures of naive conventional CD4+ T cells (Tc) in the presence of APC and anti-CD3 mAb. We found that these conditions preferentially induced conversion of Treg to IL-17 producing cells. To determine the role of IL-1β in this conversion process, we used IL-1β neutralizing antibody. Addition of anti-IL-1β neutralizing antibody reduced IL-17 production to almost undetectable levels. Because it has exogenous IL-6 can induce IL-17 production by both Treg and Tc, we evaluated whether endogenous IL-6 was involved in the conversion of Treg into IL-17 producing cells in our system. Addition of a combination of IL-6 neutralizing and IL-6 receptor blocking antibodies did not affect IL-17 production, suggesting that the conversion process of Treg into IL-17 producing cells was dependent on endogenous IL-1β rather than IL-6. To determine whether IL-1β was mandatory for this process, we used T cells from IL-1R deficient mice. Individual culture of IL−1R−/− Tc or IL-1R−/− Treg with wild type (wt) APC and co-culture of IL-1R−/− Tc and IL-1R−/− Treg with wt APC did not result in detectable IL-17 production. Similarly, no IL-17 production was observed when wt instead of IL-1R−/− Tc were used. In contrast, substitution of IL-1R−/− Treg with wt Treg resulted in abundant IL-17 production. To investigate the in vivo biological relevance of our findings we adoptively transferred Treg cells from either congenic B6.PL mice or IL-1R1−/− mice into IL-1R1−/− recipients, which were then immunized with KLH in IFA. Three days after immunization both IL-1R−/− Treg and IL-1R−/− Tc cells were incapable of producing detectable levels of IL-17 or expressing RORγt, the key transcriptional factor of IL-17. In contrast, a significant percentage of IL-17 and RORγt positive cells were detected within the adoptively transferred Thy1.1+ Treg population. Mechanistic analysis revealed that IL-1β induced activation of p38 and JNK in Treg and addition of pharmacologic inhibitors specific for these MAPKs abrogated IL-17 production. Our studies reveal that although Treg have primarily immunosuppressive functions they may also facilitate pro-inflammatory responses as they can be converted into IL-17 producing cells by IL-1β. These observations may have significant implications on clinical strategies that employ Treg for control of GvHD and suggest that further intervention might be required to prevent attainment of pro-inflammatory properties by Treg while maintaining their suppressive function. Disclosures: No relevant conflicts of interest to declare.

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Evaluation of factor VIII value in normal women and whom encountered recurrent pregnancy loss: is there any significant difference?

International Journal of Reproduction Contraception Obstetrics and Gynecology ◽

10.18203/2320-1770.ijrcog20200870 ◽

2020 ◽

Vol 9 (3) ◽

pp. 1034

Author(s):

Douaa Al Rez ◽

Hasan Naser Eldine ◽

Marwan Alhalabi

Keyword(s):

Factor Viii ◽

Pregnancy Loss ◽

Recurrent Pregnancy Loss ◽

Fetal Loss ◽

Control Group ◽

Increased Risk ◽

Significant Difference ◽

History Of ◽

Normal Women ◽

High Level

Background: Recurrent pregnancy loss (RPL) is a serious problem on the women, it defined as two or more consecutive pregnancy losses before the fetus has reached birth. The aim of this study is to evaluate the association between the elevation in the factor VIII and RPL. Because women who have thrombophilia have increased risk of fetal loss in most studies.Methods: A total 72 women were recruited in this case control study. They divided into two groups: the RPL group included 41 women with a history of recurrent pregnancy loss and the control group included 31 healthy women, who had at least one successful pregnancy and none of them had a history of fetal loss or complicated pregnancy.Results: A majority of the patients of this study didn't have a high level of factor VIII, 9 of 41 (22%) patients of RPL group in comparison with 21 of 32 (65,6%) of control group, that suffer from the increase rate of FVIII, this means that factor VIII doesn't effect on RPL.Conclusions: The present study showed that the serum elevation in the factor VIII is not significantly associated with RPL.

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Recurrent Non Immune Fetal Hydrops Associated With IPEX Syndrome

Pediatric and Developmental Pathology ◽

10.1177/1093526619834809 ◽

2019 ◽

Vol 22 (5) ◽

pp. 465-471 ◽

Cited By ~ 3

Author(s):

Elisheva Shanes ◽

Lauren Propst ◽

David W Ouyang ◽

Linda M Ernst

Keyword(s):

Pregnancy Loss ◽

Recurrent Pregnancy Loss ◽

T Regulatory Cells ◽

Novel Mutation ◽

Clinical Syndrome ◽

Fetal Anomalies ◽

Fetal Hydrops ◽

Entire Family ◽

Autoimmune Conditions ◽

Pathologic Analysis

Immune dysregulation, polyendocrinopathy, enteropathy, X-linked (IPEX) is a clinical syndrome associated with mutations in FOXP3 and consequent abnormalities of T regulatory cells. Affected males typically die in infancy or early childhood from a variety of autoimmune conditions. Reports of recurrent pregnancy loss of male fetuses in these families have been accompanied by descriptions of nonimmune fetal hydrops, with or without additional fetal anomalies. Here, we report an additional family affected by IPEX with a novel mutation leading to recurrent second trimester fetal hydrops and intrauterine fetal demise with associated fetal anomalies. This report underscores how careful genetic and pathologic analysis of even midtrimester fetuses can provide important information impacting an entire family. It also further substantiates the use of broad, symptom-targeted genetic screening panels in cases of recurrent pregnancy loss even in the absence of a remarkable pedigree.

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T-Regulatory Cells Confer Increased Myelination and Stem Cell Activity after Stroke-Induced White Matter Injury

Journal of Clinical Medicine ◽

10.3390/jcm8040537 ◽

2019 ◽

Vol 8 (4) ◽

pp. 537 ◽

Cited By ~ 8

Author(s):

Sydney Zarriello ◽

Elliot G. Neal ◽

Yuji Kaneko ◽

Cesario V. Borlongan

Keyword(s):

Cell Death ◽

Stem Cell ◽

White Matter ◽

T Regulatory Cells ◽

Cell Activity ◽

Glucose Deprivation ◽

White Matter Injury ◽

Activity Levels ◽

Regulatory Cells ◽

Treg Population

Stroke-induced hypoxia causes oligodendrocyte death due to inflammation, lack of oxygen and exacerbation of cell death. Bone marrow-derived stem cells (BMSCs) possess an endogenous population of T-regulatory cells (Tregs) which reduce secretion of pro-inflammatory cytokines that lead to secondary cell death. Here, we hypothesize that oligodendrocyte progenitor cells (OPCs) cultured with BMSCs containing their native Treg population show greater cell viability, less pro-inflammatory cytokine secretion and greater myelin production after exposure to oxygen-glucose deprivation and reoxygenation (OGD/R) than OPCs cultured without Tregs. OPCs were cultured and then exposed to OGD/R. BMSCs with or without Tregs were added to the co-culture immediately after ischemia. The Tregs were depleted by running the BMSCs through a column containing a magnetic substrate. Fibroblast growth factor beta (FGF-β) and interleukin 6 (IL-6) ELISAs determined BMSC activity levels. Immunohistochemistry assessed OPC differentiation. OPCs cultured with BMSCs containing their endogenous Tregs showed increased myelin production compared to the BMSCs with depleted Tregs. IL-6 and FGF-β were increased in the group cultured with Tregs. Collectively, these results suggest that BMSCs containing Tregs are more therapeutically active, and that Tregs have beneficial effects on OPCs subjected to ischemia. Tregs play an important role in stem cell therapy and can potentially treat white matter injury post-stroke.

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MO203ALTERATIONS IN T REGULATORY CELLS IN CHRONIC KIDNEY DISEASE PATIENTS UNDERGOING HEMODIALYSIS

Nephrology Dialysis Transplantation ◽

10.1093/ndt/gfab092.0081 ◽

2021 ◽

Vol 36 (Supplement_1) ◽

Author(s):

Erasmia Sampani ◽

Asimina Fylaktou ◽

Maria Stangou ◽

Xaralampos Vagiotas ◽

Efstratios Kasimatis ◽

...

Keyword(s):

Renal Transplantation ◽

Lymphocyte Count ◽

T Regulatory Cells ◽

Absolute Number ◽

Regulatory Cells ◽

Cell Immunity ◽

Treg Population ◽

Group A ◽

Group B

Abstract Background and Aims Disturbances in T cell immunity are frequently seen in patients with end stage renal disease (ESRD), and the effect of long term hemodialysis (HD) is still obscure, though extremely important, especially regarding T regulatory cells (Tregs, CD4+CD25+FoxP3+) which seem to have a central role in immune response and tolerance after renal transplantation. In the present study, we assessed the possible effect of HD vintage on Treg population. Method Cytometric analysis was performed in 66 patients with ESRD on HD, in order to estimate CD4+ and CD4+CD25+FoxP3+ subtypes. According to HD vintage, patients were classified into two groups, group A: recently commenced on HD, and group B: on long term HD (≤12months and >12months, respectively). Results In all 66 patients there was a negative correlation between time on HD and lymphocyte count, both percentage (r=-0.34 p=0.005) and absolute lymphocyte number (r=-0.42 p<0.001) as well as CD4+ cells frequencies (r=-0.25 p=0.042) and total numbers (r=-0.41 p=0.001). Twenty eight patients (42%) were included in group A (HD vintage 9±3 months) and 38 (58%) in group B, (HD vintage 99±41 months). Compared to group A, patients in group B, showed a significant reduction in percentage of Tregs on total lymphocytes (2.6±1.3% vs 1.9±0.9%, respectively, p<0.019) as well as absolute number of Tregs (34±14 μ/L vs. 21±11 μ/L, respectively, p<0.001). Conclusion Long term HD may act as an additional factor reducing lymphocyte count, especially Tregs in ESRD patients. This result, apart from direct affecting immunity of patients, is mostly important for those preparing for renal transplantation, or being on the waiting list.

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